Rapid and Simple Paper Strip Test for the Multiplexed Detection of Gastrointestinal Pathogens at the Point-of-Need GoDx Product Summary: This Phase 1 project will demonstrate the feasibility of developing a paper strip device for rapid (<30 minutes) and simple multiplexed detection of a 7 gastrointestinal (GI) pathogens that cause diarrheal disease. The pathogens include C. Difficile (for the U.S.) and the 6 pathogens (Cryptosporidium, Giardia lamblia, Rotavirus, Shigella, Enterotoxigenic Escherichia coli (ETEC), and Enteroaggregative Escherichia coli (EAggEC)) commonly found in the developing world. A highly innovative, simple and low cost sample preparation and concentration will be carried out using hollow silica microspheres that efficiently capture total nucleic acids and floats to the air- liquid interface for collection. The captured nucleic acids will be released onto the center of a novel star shaped lateral flow device for multiplexed detection of pathogens. Analytical validation will be conducted including assessments of reproducibility (%CV <10), sensitivity (> 90%), specificity (>90%), limit of detection (10,000 cells/g stool), and dynamic range (3 logs) on pathogen spiked-in samples or pathogen infected samples. The accuracy of the test will be benchmarked using current gold standard diagnostic methods, such as microscopy, culture, PCR and ELISA kit and the FDA-approved Luminex Gastrointestinal Pathogen Panel (GPP) bead detection, and Biofire/Biomerieux GI Panel.